The Ras proteins (Ha-Ras, Ki4a-Ras, Ki4b-Ras and N-Ras) are part of a signalling pathway that links cell surface growth factor receptors to nuclear signals initiating cellular proliferation. Biological and biochemical studies of Ras action indicate that Ras functions like a G-regulatory protein. In the inactive state, Ras is bound to GDP. Upon growth factor receptor activation Ras is induced to exchange GDP for GTP and undergoes a conformational change. The GTP-bound form of Ras propagates the growth stimulatory signal until the signal is terminated by the intrinsic GTPase activity of Ras, which returns the protein to its inactive GDP bound form (D. R. Lowy and D. M. Willumsen, Ann. Rev. Biochem. 62:851-891 (1993)). Mutated ras genes (Ha-ras, Ki4a-ras, Ki4b-ras and N-ras) are found in many human cancers, including colorectal carcinoma, exocrine pancreatic carcinoma, and myeloid leukemias. The protein products of these genes are defective in their GTPase activity and constitutively transmit a growth stimulatory signal.
Ras must be localized to the plasma membrane for both normal and oncogenic functions. At least 3 post-translational modifications are involved with Ras membrane localization, and all 3 modifications occur at the C-terminus of Ras. The Ras C-terminus contains a sequence motif termed a xe2x80x9cCAAXxe2x80x9d or xe2x80x9cCys-Aaa1-Aaa1-Aaa2-Xaaxe2x80x9d box (Cys is cysteine, Aaa is an aliphatic amino acid, the Xaa is any amino acid) (Willumsen, et al., Nature 310:583-586 (1984)). Depending on the specific sequence, this motif serves as a signal sequence for the enzymes farnesyl-protein transferase or geranylgeranyl-protein transferase, which catalyze the alkylation of the cysteine residue of the CAAX motif with a C15 or C20 isoprenoid, respectively. (S. Clarke, Ann. Rev. Biochem. 61:355-386 (1992); W. R. Schafer and J. Rine, Ann. Rev. Genetics 30:209-237 (1992)). The term prenyl-protein transferase may be used to generally refer to farnesyl-protein transferase and geranylgeranyl-protein transferase. The Ras protein is one of several proteins that are known to undergo post-translational farnesylation. Other farnesylated proteins include the Ras-related GTP-binding proteins such as Rho, fungal mating factors, the nuclear lamins, and the gamma subunit of transducin. James, et al., J. Biol. Chem. 269, 14182 (1994) have identified a peroxisome associated protein Pxf which is also farnesylated. James, et al., have also suggested that there are farnesylated proteins of unknown structure and function in addition to those listed above.
Inhibition of farnesyl-protein transferase has been shown to block the growth of Ras-transformed cells in soft agar and to modify other aspects of their transformed phenotype. It has also been demonstrated that certain inhibitors of farnesyl-protein transferase selectively block the processing of the Ras oncoprotein intracellularly (N. E. Kohl et al., Science, 260:1934-1937 (1993) and G. L. James et al., Science, 260:1937-1942 (1993). Recently, it has been shown that an inhibitor of farnesyl-protein transferase blocks the growth of ras-dependent tumors in nude mice (N. E. Kohl et al., Proc. Natl. Acad. Sci U.S.A., 91:9141-9145 (1994) and induces regression of mammary and salivary carcinomas in ras transgenic mice (N. E. Kohl et al., Nature Medicine, 1:792-797 (1995).
Indirect inhibition of farnesyl-protein transferase in vivo has been demonstrated with lovastatin (Merck and Co., Rahway, N.J.) and compactin (Hancock, et al., ibid; Casey et al., ibid; Schafer et al., Science 245:379 (1989)). These drugs inhibit HMG-CoA reductase, the rate limiting enzyme for the production of polyisoprenoids including farnesyl pyrophosphate. Farnesyl-protein transferase utilizes farnesyl pyrophosphate to covalently modify the Cys thiol group of the Ras CAAX box with a farnesyl group (Reiss et al., Cell, 62:81-88 (1990); Schaber et al., J. Biol. Chem., 265:14701-14704 (1990); Schafer et al., Science, 249:1133-1139 (1990); Manne et al., Proc. Natl. Acad. Sci USA, 87:7541-7545 (1990)). Inhibition of farnesyl pyrophosphate biosynthesis by inhibiting HMG-CoA reductase blocks Ras membrane localization in cultured cells. However, direct inhibition of farnesyl-protein transferase would be more specific and attended by fewer side effects than would occur with the required dose of a general inhibitor of isoprene biosynthesis.
Inhibitors of farnesyl-protein transferase (FPTase) have been described in two general classes. The first are analogs of farnesyl diphosphate (FPP), while the second class of inhibitors is related to the protein substrates (e.g., Ras) for the enzyme. The peptide derived inhibitors that have been described are generally cysteine containing molecules that are related to the CAAX motif that is the signal for protein prenylation. (Schaber et al., ibid; Reiss et. al., ibid; Reiss et al., PNAS, 88:732-736 (1991)). Such inhibitors may inhibit protein prenylation while serving as alternate substrates for the farnesyl-protein transferase enzyme, or may be purely competitive inhibitors (U.S. Pat. No. 5,141,851, University of Texas; N. E. Kohl et al., Science, 260:1934-1937 (1993); Graham, et al., J. Med. Chem., 37, 725 (1994)). In general, deletion of the thiol from a CAAX derivative has been shown to dramatically reduce the inhibitory potency of the compound. However, the thiol group potentially places limitations on the therapeutic application of FPTase inhibitors with respect to pharmacokinetics, pharmacodynamics and toxicity. Therefore, a functional replacement for the thiol is desirable.
It has recently been reported that farnesyl-protein transferase inhibitors are inhibitors of proliferation of vascular smooth muscle cells and are therefore useful in the prevention and therapy of arteriosclerosis and diabetic disturbance of blood vessels (JP H7-112930).
It has recently been disclosed that certain tricyclic compounds which optionally incorporate a piperidine moiety are inhibitors of FPTase (WO 95/10514, WO 95/10515 and WO 95/10516). Imidazole-containing inhibitors of farnesyl protein transferase have also been disclosed (WO 95/09001 and EP 0 675 112 A1).
It is, therefore, an object of this invention to develop peptidomimetic compounds that do not have a thiol moiety, and that will inhibit prenyl-protein transferase and thus, the post-translational prenylation of proteins. It is a further object of this invention to develop chemotherapeutic compositions containing the compounds of this invention and methods for producing the compounds of this invention.
The present invention is directed to peptidomimetic macrocyclic compounds that comprise a bicyclic imidazolyl moiety and that inhibit the prenyl-protein transferase. Further contained in this invention are chemotherapeutic compositions containing these prenyl-protein transferase inhibitors and methods for their production.
The compounds of this invention are illustrated by the formula A: 
The compounds of this invention are useful in the inhibition of prenyl-protein transferase and the prenylation of the oncogene protein Ras. In a first embodiment of this invention, the inhibitors of prenyl-protein transferase are illustrated by the formula A: 
wherein:
R1, R3, R3a and R3b are independently selected from:
a) hydrogen,
b) aryl, heterocycle, C3-C10 cycloalkyl, R10Oxe2x80x94, R11S(O)mxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2Nxe2x80x94C(O)xe2x80x94, CN, NO2, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10C(O)xe2x80x94, R10OC(O)xe2x80x94, xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94,
c) unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl or unsubstituted or substituted C2-C6 alkynyl, wherein the substituent on the substituted C1-C6 alkyl, substituted C2-C6 alkenyl or substituted C2-C6 alkynyl is selected from unsubstituted or substituted aryl, heterocyclic, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, R10Oxe2x80x94, R10S(O)mxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2Nxe2x80x94C(O)xe2x80x94, CN, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10C(O)xe2x80x94, R10OC(O)xe2x80x94, xe2x80x94N(R10)2, and R11OC(O)xe2x80x94NR10xe2x80x94,
or two R1s, two R3s, two R3as or two R3bs, on the same carbon atom may be combined to form xe2x80x94(CH2)uxe2x80x94;
R2 is independently selected from H; unsubstituted or substituted C1-8 alkyl, unsubstituted or substituted C2-8 alkenyl, unsubstituted or substituted C2-8 alkynyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocycle, 
wherein the substituted group is substituted with one or more of:
1) aryl or heterocycle, unsubstituted or substituted with:
a) C1-4 alkyl,
b) (CH2)pOR6,
c) (CH2)pNR6R7,
d) halogen,
e) CN,
2) C3-6 cycloalkyl,
3) OR6,
4) SR5, S(O)R5, SO2R5, 
two R2s attached to the same carbon atom are combined to form xe2x80x94(CH2)uxe2x80x94 wherein one of the carbon atoms is optionally replaced by a moiety selected from O, S(O)m, xe2x80x94NC(O)xe2x80x94, and xe2x80x94N(COR10)xe2x80x94;
R4 is selected from:
a) hydrogen,
b) aryl, heterocycle, C3-C10 cycloalkyl, (R10)2Nxe2x80x94C(O)xe2x80x94, (R10)2Nxe2x80x94 C(NR10)xe2x80x94, R10C(O)xe2x80x94 or R10OC(O)xe2x80x94, and
c) unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted C2-C6 alkenyl or unsubstituted or substituted C2-C6 alkynyl, wherein the substituent on the substituted C1-C6 alkyl, substituted C2-C6 alkenyl or substituted C2-C6 alkynyl is selected from unsubstituted or substituted aryl, heterocyclic, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, one or more fluorines, R10Oxe2x80x94, R11S(O)mxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2Nxe2x80x94C(O)xe2x80x94, CN, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10OC(O)xe2x80x94, R10OC(O)xe2x80x94, xe2x80x94N(R10)2, and R11OC(O)xe2x80x94NR10xe2x80x94;
R5 is selected from C1-C4 alkyl, C3-C6 cycloalkyl, heterocycle, aryl, unsubstituted or substituted with:
a) C1-4 alkoxy,
b) aryl or heterocycle,
c) halogen,
d) HO,
e) 
f) xe2x80x94SO2R11 
g) N(R10)2, or
h) one or more fluorines;
R6, R7 and R7a are independently selected from:
1) hydrogen,
2) R10OC(O)xe2x80x94, or R10OC(O)xe2x80x94, and
3) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, heterocycle, aryl, aroyl, heteroaroyl, arylsulfonyl, heteroarylsulfonyl, unsubstituted or substituted with one or more substituents selected from:
a) R10Oxe2x80x94,
b) aryl or heterocycle,
c) halogen,
d) R10C(O)NR10xe2x80x94,
e) 
f) xe2x80x94SO2R11,
g) N(R10)2,
h) C3-C6 cycloalkyl,
i) C1-C6 perfluoroalkyl,
j) (R10)2Nxe2x80x94C(NR10)xe2x80x94,
k) R10OC(O)xe2x80x94,
l) R11OC(O)NR10xe2x80x94,
m) CN, and
n) NO2; or
R6 and R7 may be joined in a ring;
R7 and R7a may be joined in a ring;
R8 is independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, unsubstituted or substituted heterocycle, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, Br, R12Oxe2x80x94, R11S(O)mxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2NC(O)xe2x80x94, R102Nxe2x80x94C(NR10)xe2x80x94, CN, NO2, R10C(O)xe2x80x94, R10OC(O)xe2x80x94, xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94, and
c) C1-C6 alkyl unsubstituted or substituted by unsubstituted or substituted aryl, unsubstituted or substituted heterocycle, C3-C10 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, Br, R10Oxe2x80x94, R11S(O)mxe2x80x94, R10C(O)NHxe2x80x94, (R10)2NC(O)xe2x80x94, R102Nxe2x80x94 C(NR10)xe2x80x94, CN, R10C(O)xe2x80x94, R10OC(O)xe2x80x94, xe2x80x94N(R10)2, or R10OC(O)NHxe2x80x94;
R9 is selected from:
a) hydrogen,
b) C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, Br, R10Oxe2x80x94, R11S(O)mxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2NC(O)xe2x80x94, R102Nxe2x80x94 C(NR10)xe2x80x94, CN, NO2, R10C(O)xe2x80x94, R10OC(O)xe2x80x94, xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94, and
c) C1-C6 alkyl unsubstituted or substituted by C1-C6 perfluoroalkyl, F, Cl, Br, R10)xe2x80x94, R11S(O)mxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2NC(O)xe2x80x94, R102Nxe2x80x94C(NR10)xe2x80x94, CN, R10C(O)xe2x80x94, R10OC(O)xe2x80x94, xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94;
R10 is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkyl substituted with one or more fluorines, benzyl, unsubstituted or substituted aryl and unsubstituted or substituted heterocycle;
R11 is independently selected from C1-C6 alkyl, C1-C6 alkyl substituted with one or more fluorines, unsubstituted or substituted aryl and unsubstituted or substituted heterocycle;
R12 is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkyl substituted with one or more fluorines, unsubstituted or substituted benzyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocycle, and C1-C6 alkyl substituted with unsubstituted or substituted aryl or unsubstituted or substituted heterocycle;
A1 is selected from a bond, xe2x80x94C(O)xe2x80x94, xe2x80x94C(O)NR10xe2x80x94, xe2x80x94NR10C(O)xe2x80x94, O, xe2x80x94N(R10)xe2x80x94, xe2x80x94S(O)2N(R10)xe2x80x94, xe2x80x94N(R10)S(O)2xe2x80x94, xe2x80x94NR10C(O)NR10xe2x80x94 and S(O)m;
A2 is selected from a bond, xe2x80x94C(O)xe2x80x94, xe2x80x94C(O)NR10xe2x80x94, xe2x80x94NR10C(O)xe2x80x94, O, xe2x80x94N(R10)xe2x80x94, xe2x80x94S(O)2N(R10)xe2x80x94, xe2x80x94N(R10)S(O)2xe2x80x94, xe2x80x94NR10C(O)NR10xe2x80x94 and S(O)m;
V is selected from:
a) heteroaryl, and
b) aryl;
W is S(O)m, O or CH2;
X is selected from: a bond, xe2x80x94C(O)xe2x80x94, xe2x80x94NR10C(O)xe2x80x94, xe2x80x94C(O)NR10xe2x80x94, xe2x80x94NR10xe2x80x94, xe2x80x94N(R10)S(O)2xe2x80x94 and S(O)2;
Y is selected from a bond, xe2x80x94C(O)xe2x80x94, xe2x80x94C(O)NR10xe2x80x94, xe2x80x94NR10C(O) xe2x80x94, xe2x80x94NR10xe2x80x94, xe2x80x94C(O)Oxe2x80x94 and xe2x80x94S(O)m;
Z1 is selected from unsubstituted or substituted aryl and unsubstituted or substituted heterocycle, wherein the substituted aryl or substituted heterocycle is substituted with one or more of:
1) C1-8 alkyl, C2-8 alkenyl or C2-8 alkynyl, unsubstituted or substituted with:
a) C1-4 alkoxy,
b) NR6R7,
c) C3-C6 cycloalkyl,
d) aryl or heterocycle,
e) HO,
f) xe2x80x94S(O)mR5,
g) xe2x80x94C(O)NR6R7, or
h) one or more fluorines;
2) substituted or unsubstituted aryl or substituted or unsubstituted heterocycle,
3) halogen,
4) OR6,
5) NR6R7,
6) CN,
7) NO2,
8) CF3;
9) xe2x80x94S(O)mR5,
10) xe2x80x94OS(O)2R5,
11) xe2x80x94C(O)NR6R7,
12) xe2x80x94C(O)OR6, or
13) C3-C6 cycloalkyl;
provided that Z1 is not selected from piperazinyl, oxopiperazinyl, dioxopiperazinyl, piperidinyl, oxopiperidinyl, pyrrolidinyl or oxopyrrolidinyl;
Z2 is selected from a bond, unsubstituted or substituted aryl and unsubstituted or substituted heteroaryl, wherein the substituted aryl or substituted heteroaryl is substituted with one or more of:
1) C1-8 alkyl, C2-8 alkenyl or C1-8 alkynyl, unsubstituted or substituted with:
a) C1-C4 alkoxy,
b) NR6R7,
c) C3-C6 cycloalkyl,
d) aryl or heterocycle,
e) HO,
f) xe2x80x94S(O)mR5,
g) xe2x80x94C(O)NR6R7, or
h) one or more fluorines;
2) substituted or unsubstituted aryl or substituted or unsubstituted heterocycle,
3) halogen,
4) OR6,
5) NR6R7,
6) CN,
7) NO2,
8) CF3;
9) xe2x80x94S(O)mR5,
10) xe2x80x94OS(O)2R5,
11) xe2x80x94C(O)NR6R7,
12) xe2x80x94C(O)OR6, or
13) C3-C6 cycloalkyl;
provided that Z2 is not selected from piperazinyl, oxopiperazinyl, dioxopiperazinyl, piperidinyl, oxopiperidinyl, pyrrolidinyl or oxopyrrolidinyl;
m is 0, 1 or 2;
n is 1, 2 or 3;
p is 0, 1, 2, 3 or 4;
q is 1 or 2;
r is 0 to 5;
s is independently 0, 1, 2 or 3;
t is 1, 2, 3 or 4; and
u is 2, 3, 4 or 5;
or a pharmaceutically acceptable salt or stereoisomer thereof.
Another embodiment of the compounds of this invention is illustrated by the formula B: 
wherein:
R1, R3 and R3b are independently selected from:
a) hydrogen,
b) R10Oxe2x80x94, xe2x80x94NR10)2, R10C(O)NR10xe2x80x94, R11OC(O)Oxe2x80x94 or R11OC(O)NR10xe2x80x94, and
c) C1-C6 alkyl, unsubstituted or substituted by R10Oxe2x80x94, xe2x80x94N(R10)2, R11C(O)NR10xe2x80x94, R11OC(O)Oxe2x80x94, R11OC(O)NR10xe2x80x94 or R11S(O)mxe2x80x94;
R2 is selected from H; 
and C1-5 alkyl, unbranched or branched, unsubstituted or substituted with one or more of:
1) aryl,
2) heterocycle,
3) OR6,
4) SR5, SO2R5, or
5) 
R4 is selected from:
a) hydrogen, and
b) unsubstituted or substituted C1-C6 alkyl, wherein the substituent on the substituted C1-C6 alkyl is selected from one or more fluorines, R10Oxe2x80x94, R 11S(O)mxe2x80x94, R10C(O)NR10xe2x80x94, R10OC(O)Oxe2x80x94 and R11OC(O)xe2x80x94 NR10;
R5 is selected from:
C1-4 alkyl and C3-C6 cycloalkyl, unsubstituted or substituted with:
a) C1-4 alkoxy,
b) one or more fluorines, or
c) aryl or heterocycle;
R6 and R7 are independently selected from:
a) hydrogen,
b) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, R10C(O)xe2x80x94 or R10OC(O)xe2x80x94 and
c) C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R10Oxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10C(O)xe2x80x94, R10OC(O)xe2x80x94, xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94;
R8 is independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R10Oxe2x80x94, R10C(O)NR10xe2x80x94, CN, NO2, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10C(O)xe2x80x94, xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94, and
c) C1-C6 alkyl substituted by unsubstituted or substituted aryl, C1-C6 perfluoroalkyl, R10Oxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10C(O)xe2x80x94, xe2x80x94N(R10)2, or R10C(O)NR10xe2x80x94;
R10 is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkyl substituted with one or more fluorines, benzyl and unsubstituted or substituted aryl;
R11 is independently selected from C1-C6 alkyl, C1-C6 alkyl substituted with one or more fluorines, and unsubstituted or substituted aryl;
R12 is independently selected from hydrogen, C1-C6 alkyl, unsubstituted or substituted benzyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocycle, and C1-C6 alkyl substituted with one or more fluorines, unsubstituted or substituted aryl or unsubstituted or substituted heterocycle;
A1 is selected from a bond, xe2x80x94N(R10)xe2x80x94, S(O)m and O;
W is S(O)m or CH2;
X is selected from: a bond, xe2x80x94C(O)xe2x80x94, xe2x80x94NR10C(O)xe2x80x94, xe2x80x94N(R10)S(O)2xe2x80x94 and S(O)2;
Y is selected from a bond, xe2x80x94C(O)xe2x80x94, xe2x80x94C(O)NR10xe2x80x94, xe2x80x94C(O)Oxe2x80x94, xe2x80x94(CR1c2)xe2x80x94 and xe2x80x94S(O)m;
Z1 is selected from unsubstituted or substituted aryl or unsubstituted or substituted heteroaryl, wherein the substituted aryl or substituted heteroaryl is substituted with one or two of:
1) C1-8 alkyl, C2-8 alkenyl or C2-8 alkynyl, unsubstituted or substituted with:
a) C1-4 alkoxy,
b) NR6R7,
c) C3-6 cycloalkyl,
d) aryl or heterocycle,
e) HO,
f) xe2x80x94S(O)mR5,
g) xe2x80x94C(O)NR6R7, or
h) one or more fluorines;
2) substituted or unsubstituted aryl or substituted or unsubstituted heterocycle,
3) halogen,
4) OR6,
5) NR6R7,
6) CN,
7) NO2,
8) CF3,
9) xe2x80x94S(O)mR5,
10) xe2x80x94OS(O)2R5,
11) xe2x80x94C(O)NR6R7,
12) xe2x80x94C(O)OR6, or
13) C3-C6 cycloalkyl;
provided that Z1 is not selected from piperazinyl, oxopiperazinyl, dioxopiperazinyl, piperidinyl, oxopiperidinyl, pyrrolidinyl or oxopyrrolidinyl;
m is 0, 1 or 2;
p is 0, 1, 2, 3 or 4;
q is 1 or 2;
r is 0 to 5;
s is independently 0, 1, 2 or 3; and
t is 1, 2, 3 or 4;
or a pharmaceutically acceptable salt or stereoisomer thereof.
In another embodiment of this invention, the inhibitors of prenyl-protein transferase are illustrated by the formula C: 
wherein:
R1, R3 and R3b are independently selected from:
a) hydrogen,
b) R10Oxe2x80x94, N(R10)2, R10C(O)NR10xe2x80x94, R11OC(O)Oxe2x80x94 or R11OC(O)NR10xe2x80x94, and
c) C1-C6 alkyl, unsubstituted or substituted R10Oxe2x80x94, N(R10)2, R10C(O)NR10xe2x80x94, R11OC(O)Oxe2x80x94, R11OC(O)NR10xe2x80x94 or R11S(O)mxe2x80x94;
R2 is selected from H; 
and C1-5 alkyl, unbranched or branched, unsubstituted or substituted with one or more of:
1) aryl,
2) heterocycle,
3) OR6,
4) SR5, SO2R5, or
5) 
R4 is selected from:
a) hydrogen, and
b) unsubstituted or substituted C1-C6 alkyl, wherein the substituent on the substituted C1-C6 alkyl is selected from one or more fluorines, R10Oxe2x80x94, R 11S(O)mxe2x80x94, R10C(O)NR10xe2x80x94, R10OC(O)Oxe2x80x94 and R11OC(O)xe2x80x94 NR10;
R5 is selected from C1-4 alkyl and C3-C6 cycloalkyl, unsubstituted or substituted with:
a) C1-C4 alkoxy,
b) halogen, or
c) aryl or heterocycle;
R6 and R7 are independently selected from:
a) hydrogen,
b) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, R10C(O)xe2x80x94 or R10OC(O)xe2x80x94 and
c) C1-C6 alkyl substituted by C1-C6 perfluoroalkyl, R10Oxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10C(O)xe2x80x94, R10C(O), xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94;
R8 is independently selected from:
a) hydrogen,
b) unsubstituted or substituted aryl, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl, F, Cl, R12Oxe2x80x94, R10C(O)NR10xe2x80x94, CN, NO2, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10C(O)xe2x80x94, xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94, and
c) C1-C6 alkyl substituted by unsubstituted or substituted aryl, C1-C6 perfluoroalkyl, R10Oxe2x80x94, R10C(O)NR10xe2x80x94, (R10)2Nxe2x80x94C(NR10)xe2x80x94, R10C(O)xe2x80x94, xe2x80x94N(R10)2, or R11OC(O)NR10xe2x80x94;
R10 is independently selected from hydrogen, C1-C6 alkyl, C1-C6 alkyl substituted with one or more fluorines, benzyl and unsubstituted or substituted aryl;
R11 is independently selected from C1-C6 alkyl, C1-C6 alkyl substituted with one or more fluorines and unsubstituted or substituted aryl;
R12 is independently selected from hydrogen, C1-C6 alkyl, unsubstituted or substituted benzyl, unsubstituted or substituted aryl, unsubstituted or substituted heterocycle, and C1-C6 alkyl substituted with one or more fluorines, unsubstituted or substituted aryl or unsubstituted or substituted heterocycle;
A1 is selected from a bond, xe2x80x94N(R10)xe2x80x94, S(O)m and O;
W is S(O)m or CH2;
X is selected from: xe2x80x94C(O)xe2x80x94, xe2x80x94C(O)NR10xe2x80x94, xe2x80x94N(R10)xe2x80x94, xe2x80x94S(O)2N(R10)xe2x80x94 and S(O)m;
Y is selected from: a bond, xe2x80x94C(O)xe2x80x94, xe2x80x94NR10C(O)xe2x80x94, xe2x80x94N(R10)xe2x80x94, xe2x80x94N(R10)S(O)2xe2x80x94, and S(O)m;
Z1 is selected from unsubstituted or substituted aryl or unsubstituted or substituted heteroaryl, wherein the substituted aryl or substituted heteroaryl is substituted with one or two of:
1) C1-8 alkyl, C2-8 alkenyl or C1-8 alkynyl, unsubstituted or substituted with:
a) C1-C4 alkoxy,
b) NR6R7,
c) C3-C6 cycloalkyl,
d) aryl or heterocycle,
e) HO,
f) xe2x80x94S(O)mR5,
g) xe2x80x94C(O)NR6R7, or
h) one or more fluorines;
2) substituted or unsubstituted aryl or substituted or unsubstituted heterocycle,
3) halogen,
4) OR6,
5) NR6R7,
6) CN,
7) NO2,
8) CF3,
9) xe2x80x94S(O)mR5,
10) xe2x80x94OS(O)2R5,
11) xe2x80x94C(O)NR6R7,
12) xe2x80x94C(O)OR6, or
13) C3-C6 cycloalkyl;
provided that Z1 is not selected from piperazinyl, oxopiperazinyl, dioxopiperazinyl, piperidinyl, oxopiperidinyl, pyrrolidinyl or oxopyrrolidinyl;
m is 0, 1 or 2;
p is 0, 1, 2, 3 or 4;
q is 1 or 2;
r is 0 to 5;
s is independently 0, 1, 2 or 3; and
t is 1, 2, 3 or 4;
or a pharmaceutically acceptable salt or stereoisomer thereof.
Examples of the compounds of the invention are: 
(xc2x1)-15-Bromo-18,19,20,21-tetrahydro-19,22-dioxo-26-thia-5H,17H-3,5-ethano-6,10:12,16-dimetheno-22H-imidazo[3,4-h][1,8,11,14] oxatriazacycloeicosine-9-carbonitrile 
(xc2x1)-15-Bromo-18,19,20,21-tetrahydro-22-oxo-26-thia-5H,17H-3,5-ethano-6,10:12,16-dimetheno-22H-imidazo[3,4-h][1,8,11,14] oxatriazacycloeicosine-9-carbonitrile 
(xc2x1)-15-Bromo-18,19,20,21-tetrahydro-22-oxo-18,26-dithia-5H, 17H-3,5-ethano-6,10:12,16-dimetheno-22H-imidazo[3,4-h][1,11,14] oxadiazacycloeicosine-9-carbonitrile 
(xc2x1)-15-Bromo-18,19,20,21tetrahydro-18,18,22-trioxo-18,26-dithia-5H, 17H-3,5-ethano-6,10:12,16-dimetheno-22H-imidazo[3 ,4-h][1,11,14] oxadiazacycloeicosine-9-carbonitrile 
(xc2x1)-19,20,21,22-Tetrahydro-14,14,22-trioxo-5H, 17H-3,5-ethano-6,10:12,16-dimetheno-imidazo[3,4-h][1,7,11,14]oxadiazathiacycloeicosine-9-carbonitrile 
(xc2x1)-19,20,21,22-Tetrahydro-22-oxo-5H,17H-3,5-ethano-6,10:12,16-dimetheno-imidazo[3,4-h][1,8,11 ]oxadiazacycloeicosine-9-carbonitrile
or the pharmaceutically acceptable salts or stereoisomers thereof.
The compounds of the present invention may have asymmetric centers, chiral axes and chiral planes, and occur as racemates, racemic mixtures, and as individual diastereomers, with all possible isomers, including optical isomers, being included in the present invention. (See E. L. Eliel and S. H. Wilen Stereochemistry of Carbon Compounds (John Wiley and Sons, New York 1994), in particular pages 1119-1190) When any variable (e.g. aryl, heterocycle, R1, R6 etc.) occurs more than one time in any constituent, its definition on each occurrence is independent at every other occurrence. Also, combinations of substituents/or variables are permissible only if such combinations result in stable compounds.
As used herein, xe2x80x9calkylxe2x80x9d is intended to include both branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms; xe2x80x9caloxyxe2x80x9d represents an alkyl group of indicated number of carbon atoms attached through an oxygen bridge. xe2x80x9cHalogenxe2x80x9d or xe2x80x9chaloxe2x80x9d as used herein means fluoro, chloro, bromo and iodo.
Preferably, alkenyl is C2-C6 alkenyl.
Preferably, alkynyl is C2-C6 alkynyl.
As used herein, xe2x80x9ccycloalkylxe2x80x9d is intended to include cyclic saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. Preferably, cycloalkyl is C3-C10 cycloalkyl. Examples of such cycloalkyl elements include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
As used herein, xe2x80x9carylxe2x80x9d is intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 members in each ring, wherein at least one ring is aromatic. Examples of such aryl elements include phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthryl, anthryl or acenaphthyl.
The term heterocycle or heterocyclic, as used herein, represents a stable 5- to 7-membered monocyclic or stable 8- to 11-membered bicyclic heterocyclic ring which is either saturated or unsaturated, and which consists of carbon atoms and from one to four heteroatoms selected from the group consisting of N, O, and S, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring. The heterocyclic ring may be attached at any heteroatom or carbon atom which results in the creation of a stable structure. The term heterocycle or heterocyclic, as used herein, includes heteroaryl moieties. Examples of such heterocyclic elements include, but are not limited to, azepinyl, benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, 1,3-dioxolanyl, furyl, imidazolidinyl, imidazolinyl, imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, isothiazolidinyl, morpholinyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, 2-oxopiperazinyl, 2-oxopiperdinyl, 2-oxopyrrolidinyl, piperidyl, piperazinyl, pyridyl, pyrazinyl, pyrazolidinyl, pyrazolyl, pyridazinyl, 2-pyridinonyl, pyrimidinyl, pyrrolidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiazolyl, thiazolinyl, thienofuryl, thienothienyl, thienyl and triazolyl.
As used herein, xe2x80x9cheteroarylxe2x80x9d is intended to mean any stable monocyclic or bicyclic carbon ring of up to 7 members in each ring, wherein at least one ring is aromatic and wherein from one to four carbon atoms are replaced by heteroatoms selected from the group consisting of N, O, and S. Examples of such heterocyclic elements include, but are not limited to, benzimidazolyl, benzisoxazolyl, benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl, dihydrobenzofuryl, dihydrobenzothienyl, dihydrobenzothiopyranyl, dihydrobenzothiopyranyl sulfone, furyl, imidazolyl, indolinyl, indolyl, isochromanyl, isoindolinyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxadiazolyl, pyridyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiazolyl, thienofuryl, thienothienyl, thienyl and triazolyl.
As used herein, unless otherwise specifically defined, substituted alkyl, substituted cycloalkyl, substituted aroyl, substituted aryl, substituted heteroaroyl, substituted heteroaryl, substituted arylsulfonyl, substituted heteroaryl-sulfonyl and substituted heterocycle include moieties containing from 1 to 3 substituents in addition to the point of attachment to the rest of the compound. Preferably, such substituents are selected from the group which includes but is not limited to F, Cl, Br, CF3, NH2, N(C1-C6 alkyl)2, NO2, CN, (C1-C6 alkyl)Oxe2x80x94, (aryl)Oxe2x80x94, xe2x80x94OH, (C1-C6 alkyl)S(O)mxe2x80x94, (C1-C6 alkyl)C(O)NHxe2x80x94, H2Nxe2x80x94C(NH)xe2x80x94, (C1-C6 alkyl)C(O)xe2x80x94, (C1-C6 alkyl)OC(O)xe2x80x94, (C1-C6 alkyl)OC(O)NHxe2x80x94, phenyl, pyridyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thienyl, furyl, isothiazolyl and C1-C20 alkyl.
As used herein, the term xe2x80x9cone or more fluorinesxe2x80x9d describes substitution on one or more carbon atoms of a substituted group with one or more fluroine atoms. Preferably the substituted group which is substituted with one or more fluorines is substitued with one to five fluorines. Preferably a C1-6 alkyl substituted with one or more fluorines is a C1-6 alkyl substituted with one to five fluorines.
Preferably, as used herein in the definition of R6 and R7, the substituted C1-6 alkyl, substituted C2-6 alkenyl, substituted C2-6 alkynyl, substituted C3-C6 cycloalkyl, substituted aroyl, substituted aryl, substituted heteroaroyl, substituted arylsulfonyl, substituted heteroarylsulfonyl and substituted heterocycle, include moieties containing from 1 to 3 substituents in addition to the point of attachment to the rest of the compound.
The moiety formed when, in the definition of R1, R2, R3, R3a and R3b, two R1s, two R2s, two R3s, two R3as or two R3bb, on the same carbon atom are combined to form xe2x80x94(CH2)uxe2x80x94 is illustrated by the following: 
In addition, such cyclic moieties may optionally include a heteroatom(s). Examples of such heteroatom-containing cyclic moieties include, but are not limited to: 
The moiety formed when, in the definition of R6, R7 and R7a, R6 and R7 or R7 and R7a are joined to form a ring, is illustrated by, but not limited to, the following: 
Lines drawn into the ring systems from substituents (such as from R8, R9 etc.) indicate that the indicated bond may be attached to any of the substitutable ring carbon and nitrogen atoms.
Preferably, R1, R3, R3a and R3b, are independently selected from: hydrogen, xe2x80x94N(R10)2, R10C(O)NR10xe2x80x94 or unsubstituted or substituted C1-C6 alkyl wherein the substituent on the substituted C1-C6 alkyl is selected from unsubstituted or substituted phenyl, xe2x80x94N(R10)2, R10Oxe2x80x94 and R10C(O)NR10xe2x80x94. More preferably R1, R3, R3a and R3b are hydrogen.
Preferably, R2 is selected from H, 
and an unsubstituted or substituted C1-8 alkyl,
wherein the substituted C1-8 alkyl is substituted with one or more of:
1) aryl or heterocycle, unsubstituted or substituted with:
a) C1-C4 alkyl,
b) (CH2)pOR6,
c) (CH2)pNR6R7,
d) halogen,
2) C3-C6 cycloalkyl,
3) OR6 
4) SR5, S(O)R5, SO2R5,
5) xe2x80x94NR6R7
Preferably, R4 is selected from:
a) hydrogen, and
b) unsubstituted or substituted C1-C6 alkyl, wherein the substituent on the substituted C1-C6 alkyl is selected from one or more fluorines, R10Oxe2x80x94, R11S(O)mxe2x80x94, R10C(O))NR10xe2x80x94, R10OC(O)Oxe2x80x94 and R11C(O)xe2x80x94 NR10.
Preferably, R5 is C1-C6 alkyl.
Preferably, R6, R7 and R7a is selected from: hydrogen, unsubstituted or substituted C1-C6 alkyl, unsubstituted or substituted aryl and unsubstituted or substituted cycloalkyl.
Preferably, R9 is hydrogen or methyl.
Preferably, R10 is selected from H, C1-C6 alkyl and benzyl.
Preferably, A1 is selected from a bond, O, xe2x80x94N(R10)xe2x80x94 and S(O)m.
Preferably, A2 is selected from: a bond, xe2x80x94C(O)NR10xe2x80x94, xe2x80x94NR10C(O)xe2x80x94, O, xe2x80x94N(R10)xe2x80x94, xe2x80x94S(O)2N(R10)xe2x80x94, S(xe2x95x90O)m and xe2x80x94N(R10)S(O)2xe2x80x94.
Preferably, V is selected from heteroaryl and aryl. More preferably, V is phenyl or pyridyl.
Preferably, W is S or CH2.
Preferably, X is selected from xe2x80x94NR10C(O)xe2x80x94, O, xe2x80x94N(R10)xe2x80x94 and xe2x80x94N(R10)S(O)2xe2x80x94.
Preferably, Y is selected from xe2x80x94C(O)NR10xe2x80x94, O, xe2x80x94N(R10)xe2x80x94, and xe2x80x94S(O)2N(R10)xe2x80x94.
Preferably, Z1 and Z2 are independently selected from unsubstituted or substituted phenyl, unsubstituted or substituted naphthyl, unsubstituted or substituted pyridyl, unsubstituted or substituted furanyl and unsubstituted or substituted thienyl. More preferably, Z1 is selected from unsubstituted or substituted phenyl and unsubstituted or substituted naphthyl. More preferably, Z2 is selected from a bond and unsubstituted or substituted phenyl.
Preferably, r is 1 or 2.
Preferably p is 0, 1 or 2.
Preferably s is 0 or 1.
Preferably, the moiety 
is selected from: 
It is intended that the definition of any substituent or variable (e.g., R1a, R9, n, etc.) at a particular location in a molecule be independent of its definitions elsewhere in that molecule. Thus, xe2x80x94N(R10)2 represents xe2x80x94NHH, xe2x80x94NHCH3, xe2x80x94NHC2H5, etc. It is understood that substituents and substitution patterns on the compounds of the instant invention can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art, as well as those methods set forth below, from readily available starting materials.
The pharmaceutically acceptable salts of the compounds of this invention include the conventional non-toxic salts of the compounds of this invention as formed, e.g., from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like: and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, trifluoroacetic and the like.
The pharmaceutically acceptable salts of the compounds of this invention can be synthesized from the compounds of this invention which contain a basic moiety by conventional chemical methods. Generally, the salts are prepared either by ion exchange chromatography or by reacting the free base with stoichiometric amounts or with an excess of the desired salt-forming inorganic or organic acid in a suitable solvent or various combinations of solvents.
Reactions used to generate the compounds of this invention are prepared by employing reactions as shown in the Schemes 1-13, in addition to other standard manipulations such as ester hydrolysis, cleavage of protecting groups, etc., as may be known in the literature or exemplified in the experimental procedures. Substituents Rsub and Rsubxe2x80x2, as shown in the Schemes, represent the substituents on Z1 and Z2 and other moieties of the instant compounds; however their point of attachment to the ring is illustrative only and is not meant to be limiting. It is understood that one of ordinary skill in the art would be readily able to substitute commercially available or readily prepared suitably substituted aromatic moieties for those unsubstituted moieties illustrated in the schemes.
These reactions may be employed in a linear sequence to provide the compounds of the invention or they may be used to synthesize fragments which are subsequently joined by the alkylation reactions described in the Schemes.
Synopsis of Schemes 1-13
The requisite intermediates are in some cases commercially available, or can be prepared according to literature procedures and synthetic methods well known in the medicinal chemistry art. In Scheme 1, for example, the synthesis of a key intermediate in the preparation of bicyclic imidazolyl compounds of the instant invention and its incorporation into the macrocycle is generally outlined. An (xcex1-bromoacetophenone I (commercially available, or prepared by standard procedures) is reacted with 2-thio imidazole II under basic conditions, to give thio ether III. Reduction of the ketone provides intermediate hydroxy imidazole IV. Subsequent protection of the hydroxy imidazole IV with di-tert-butyl dicarbonate gives an intermediate N-t-butoxycarbonyl imidazole which is not isolated; rather it is treated in situ with methane sulfonic acid anhydride and an amine base to mesylate the hydroxyl group. Heating this intermediate gives dihydroimidazo[2,1-b]thiazole V, the product of intramolecular alkylation, with subsequent loss of the t-butyloxycarbonyl protecting group occurring during a standard aqueous workup. Ester group saponification gives carboxylic acid intermediate VI. The ester moiety of intermediate V may alternatively reduced to the hydroxymethyl intermediate VII, which may itself be oxidized to the aldehyde intermediate VIII.
Scheme 2 illustrates preparation of a compound of the instant invention that incorporates the dihydroimidazo[2,1-b]thiazole moiety. Thus, a protected aminomethylphenol IX is reacted with a suitably substituted amino acid X to provide, after removal of the nitrogen protecting group, the intermediate XI. Intermediate XI is then coupled to the carboxylic acid intermediate VI described above to provide the bis-amide intermediate XII. Potassium fluoride mediated intramolecular cyclization provides the compound of the instant invention XIII.
Scheme 3 illustrates preparation of compounds of the instant invention wherein xe2x80x9cXxe2x80x9d is an amine moiety and xe2x80x9cYxe2x80x9d is an amide moiety. Thus the nitrogen-protected amino phenol XIV is reacted with a suitably substituted chloroacetyl chloride to provide the intermediate XV. Intermediate XV is reacted with the amine XVI, prepared as illustrated from key intermediate aldehyde VIII, to provide intermediate XVII. Potassium fluoride mediated intramolecular cyclization, followed by acid removal of the nitrogen protecting group provides the instant compound XVIII.
Preparation of compounds of the instant invention wherein xe2x80x9cWxe2x80x9d is an alkylene chain is illustrated in Scheme 3. Thus, a 1-benzyl-5-hydroxymethylimidazole XIX, prepared according to the general procedure outlined in Anthony et al., J. Med. Chem. 1999, 42, 3356-3368, is protected as the t-butyldimethylsilyl ether XX. Generation of the benzylic carbanion with a strong base such as lithium bis(trimethylsilyl)amide, and subsequent reaction with a suitable alkylating agent gives XXI. Deprotection of the t-butyldimethylsilyl ether gives primary alcohol XXII, which is converted to amine XXIII as shown. Intermediate XXIII is reacted with a suitably substituted N-naphthyl chloro acetamide XXIV, to provide intermediate XXV. The hydroxyl moiety on the naphthyl is protected and the protecting group was removed from the primary alcohol to provide intermediate XXVI. The primary alcohol was oxidized to the aldehyde XXVII and cyclization via a modified intramolecular Prins reaction yields the tetrahydroimidazo[1,2-a]pyridine or homolog intermediate XXVIII. Deoxygenation via the thiocarbonate XXIX with tri-n-butyltin hydride and 2,2xe2x80x2-azobisisobutyronitrile gives intermediate XXX. Removal of the benzyl moiety by catalytic reduction, followed by cesium carbonate mediated cyclization provides the compound of the instant invention XXXI.
Preparation of compounds of the instant invention which incorporate a urea moiety in the macrocyclic ring is illustrated in Scheme 5.
Scheme 6 illustrates the preparation of other key intermediates in the synthesis of the instant compounds. Thus, imidazolyl-2-aldehyde XXXII is first nitrogen protected and then treated with the lithium salt of a suitably substituted acetophenone XXXIII to provide the hydroxy ketone XXXIV. Dehydration of intermediate XXXIV, followed by reduction of the ketone and hydrogenation of the olefin provides alcohol XXXV. Activation of the hydroxyl moiety and heating provides the bicyclic intermediate XXXVI. The N-protecting group is removed and the intermediate treated with formaldehyde to provide the hydroxymethyl intermediate XXXVII. Intermediate XXXVII may be oxidized to the coresponding aldehyde XXXVIII, which may be further oxidized to the corresponding acid IXL Scheme 6 further illustrates preparation of the instant compound XL from the aldehyde XXXVIII.
Preparation of compounds of the instant invention wherein both xe2x80x9cXxe2x80x9d and xe2x80x9cYxe2x80x9d are a bond is illustrated in Scheme 7. Thus the aldehyde XXXVIII undergoes a Wittig reaction with a suitably subsituted phenolic alkyl Wittig reagent to provide the olefin XLI. Catalytic hydrogenation reduces the olefin and removed the benzyl group to provide intermediate XLII. Cesium carbonate mediated cyclization provides the compound of the instant invention XLIII.
Similar chemistry may also be exploited to provide compounds of the instant invention wherein sulfur is incorporated into the linker. Thus, the aldehyde XXXVIII is treated with a methylalkanoate Wittig reagent to provide the unsaturated ester XLIV. Catalytic hydrogentation, followed by reduction of the ester to the corresponding alcohol provides intermediate XLV. Intermediate XLV may then undergo a Mitsunobu reaction with thioacetic acid to provide, after removal of the acetyl group, the mercaptan XLVI. Intermediate XLVI may be treated with a suitably protected halomethylphenol to provide intermediate XLVII. Removal of the hydroxyl protecting group, followed by cesium carbonate mediated cyclization provides the compound of the instant invention XLVIII. Stoicheometric oxidation of the sulfur moiety provides either the sulfone or sulfoxide IL As shown in Scheme 9, the unsaturated ester intermediate XL may be converted to the acid L, which may then be esterified with a suitably substituted aralkyl or heteroaralkyl alcohol to provide the intermediate LI. Deprotection and cyclization as described above provides the instant compound LII. Acid intermediate L may alternatively be converted to a suitably substituted compound of the instant invention comprising a ketone moiety in the linker as illustrated in Scheme 10.
Incorporation of a sulfur containing moiety for A1 in the instant compounds is illustrated in Scheme 11. Thus chloroacetyl chloride is reacted with the tritylsulfide aniline LIII to provide the intermediate LIV. Intermediate LIV may be reacted with the previously described amine XXIII to provide intermediate LIV. Subsequent cyclization to form the fused ring, followed by removal of the hydroxy group as described above gives intermediate LV. Removal of the trityl protecting group liberates the mercaptan moiety, which undergoes cyclization under cesium carbonate conditions to provide the instant compound LVI. The sulfur may be oxidized to either the sulfone or sulfoxide LVII.
Scheme 12 illustrates the synthetic strategy that is employed when the R8 substituent is not an electronic withdrawing moiety either ortho or para to the fluorine atom. In the absence of the electronic withdrawing moiety, the intramolecular cyclization can be accomplished via an Ullmann reaction. Thus, a suitably substituted iodo benzaldehyde LVIII may be employed in place of intermediate I to provide the intermediate LIX. The previously described elaboration of the intermediate LIX provides the carboxylic acid LX. Carboxylic acid LX is then reacted with the amine XI described above to provide intermediate LXI. Desilylation followed by intramolecular cyclization under Ullmann conditions, provides the instant compound LXII.
Scheme 13 illustrates the preparation of intermediates LXIII and LXIV which may be incorporated into synthetic reactions described above to provide compounds of the instant invention wherein W is oxygen (O). 
In a preferred embodiment of the instant invention the compounds of the invention are selective inhibitors of farnesyl-protein transferase. A compound is considered a selective inhibitor of farnesyl-protein transferase, for example, when its in vitro farnesyl-protein transferase inhibitory activity, as assessed by the assay described in Example 7, is at least 100 times greater than the in vitro activity of the same compound against geranylgeranyl-protein transferase-type I in the assay described in Example 8. Preferably, a selective compound exhibits at least 1000 times greater activity against one of the enzymatic activities when comparing geranylgeranyl-protein transferase-type I inhibition and farnesyl-protein transferase inhibition.
It is also preferred that the selective inhibitor of farnesyl-protein transferase is further characterized by:
a) an IC50 (a measure of in vitro inhibitory activity) for inhibition of the prenylation of newly synthesized K-Ras protein more than about 100-fold higher than the EC50 for the inhibition of the farnesylation of hDJ protein.
When measuring such IC50s and EC50s the assays described in Example 12 may be utilized.
It is also preferred that the selective inhibitor of farnesyl-protein transferase is further characterized by:
b) an IC50 (a measurement of in vitro inhibitory activity) for inhibition of K4B-Ras dependent activation of MAP kinases in cells at least 100-fold greater than the IC50 for inhibition of the farnesylation of the protein hDJ in cells.
It is also preferred that the selective inhibitor of farnesyl-protein transferase is further characterized by:
c) an IC50 (a measurement of in vitro inhibitory activity) against H-Ras dependent activation of MAP kinases in cells at least 1000 fold lower than the inhibitory activity (IC50) against H-ras-CVLL (SEQ.ID.NO.: 1) dependent activation of MAP kinases in cells.
When measuring Ras dependent activation of MAP kinases in cells the assays described in Example 11 may be utilized.
In another preferred embodiment of the instant invention the compounds of the invention are dual inhibitors of famesyl-protein transferase and geranylgeranyl-protein transferase type I. Such a dual inhibitor may be termed a Class II prenyl-protein transferase inhibitor and will exhibit certain characteristics when assessed in in vitro assays, which are dependent on the type of assay employed.
In a SEAP assay, such as described in Examples 11, it is preferred that the dual inhibitor compound has an in vitro inhibitory activity (IC50) that is less than about 12 xcexcM against K4B-Ras dependent activation of MAP kinases in cells.
The Class II prenyl-protein transferase inhibitor may also be characterized by:
a) an IC50 (a measurement of in vitro inhibitory activity) for inhibiting K4B-Ras dependent activation of MAP kinases in cells between 0.1 and 100 times the IC50 for inhibiting the farnesylation of the protein hDJ in cells; and
b) an IC50 (a measurement of in vitro inhibitory activity) for inhibiting K4B-Ras dependent activation of MAP kinases in cells greater than 5-fold lower than the inhibitory activity (IC50) against expression of the SEAP protein in cells transfected with the pCMV-SEAP plasmid that constitutively expresses the SEAP protein.
The Class II prenyl-protein transferase inhibitor may also be characterized by:
a) an IC50 (a measurement of in vitro inhibitory activity) against H-Ras dependent activation of MAP kinases in cells greater than 2 fold lower but less than 20,000 fold lower than the inhibitory activity (IC50) against H-ras-CVLL (SEQ.ID.NO.: 1) dependent activation of MAP kinases in cells; and
b) an IC50 (a measurement of in vitro inhibitory activity) against H-ras-CVLL dependent activation of MAP kinases in cells greater than 5-fold lower than the inhibitory activity (IC50) against expression of the SEAP protein in cells transfected with the pCMV-SEAP plasmid that constitutively expresses the SEAP protein.
The Class II prenyl-protein transferase inhibitor may also be characterized by:
a) an IC50 (a measurement of in vitro inhibitory activity) against H-Ras dependent activation of MAP kinases in cells greater than 10-fold lower but less than 2,500 fold lower than the inhibitory activity (IC50) against H-ras-CVLL (SEQ.ID.NO.: 1) dependent activation of MAP kinases in cells; and
b) an IC50 (a measurement of in vitro inhibitory activity) against H-ras-CVLL dependent activation of MAP kinases in cells greater than 5 fold lower than the inhibitory activity (IC50) against expression of the SEAP protein in cells transfected with the pCMV-SEAP plasmid that constitutively expresses the SEAP protein.
A method for measuring the activity of the inhibitors of prenyl-protein transferase, as well as the instant combination compositions, utilized in the instant methods against Ras dependent activation of MAP kinases in cells is described in Example 11.
In yet another embodiment, a compound of the instant invention may be a more potent inhibitor of geranylgeranyl-protein transferase-type I than it is an inhibitor of farnesyl-protein transferase.
The instant compounds are useful as pharmaceutical agents for mammals, especially for humans. These compounds may be administered to patients for use in the treatment of cancer. Examples of the type of cancer which may be treated with the compounds of this invention include, but are not limited to, colorectal carcinoma, exocrine pancreatic carcinoma, myeloid leukemias and neurological tumors. Such tumors may arise by mutations in the ras genes themselves, mutations in the proteins that can regulate Ras activity (i.e., neurofibromin (NF-1), neu, src, abl, Ick, fyn) or by other mechanisms.
The compounds of the instant invention inhibit farnesyl-protein transferase and the farnesylation of the oncogene protein Ras. The instant compounds may also inhibit tumor angiogenesis, thereby affecting the growth of tumors (J. Rak et al. Cancer Research, 55:4575-4580 (1995)). Such anti-angiogenesis properties of the instant compounds may also be useful in the treatment of certain forms of vision deficit related to retinal vascularization.
The compounds of this invention are also useful for inhibiting other proliferative diseases, both benign and malignant, wherein Ras proteins are aberrantly activated as a result of oncogenic mutation in other genes (i.e., the Ras gene itself is not activated by mutation to an oncogenic form) with said inhibition being accomplished by the administration of an effective amount of the compounds of the invention to a mammal in need of such treatment. For example, the composition is useful in the treatment of neurofibromatosis, which is a benign proliferative disorder.
The instant compounds may also be useful in the treatment of certain viral infections, in particular in the treatment of hepatitis delta and related viruses (J. S. Glenn et al. Science, 256:1331-1333 (1992).
The compounds of the instant invention are also useful in the prevention of restenosis after percutaneous transluminal coronary angioplasty by inhibiting neointimal formation (C. Indolfi et al. Nature medicine, 1:541-545(1995).
The instant compounds may also be useful in the treatment and prevention of polycystic kidney disease (D. L. Schaffner et al. American Journal of Pathology, 142:1051-1060 (1993) and B. Cowley, Jr. et al. FASEB Journal, 2:A3160 (1988)).
The instant compounds may also be useful for the treatment of fungal infections.
The instant compounds may also be useful as inhibitors of proliferation of vascular smooth muscle cells and therefore useful in the prevention and therapy of arteriosclerosis and diabetic vascular pathologies.
The compounds of the instant invention may also be useful in the prevention and treatment of endometriosis, uterine fibroids, dysfunctional uterine bleeding and endometrial hyperplasia.
In such methods of prevention and treatment as described herein, the prenyl-protein transferase inhibitors of the instant invention may also be co-administered with other well known therapeutic agents that are selected for their particular usefulness against the condition that is being treated. For example, the prenyl-protein transferase inhibitor may be useful in further combination with drugs known to supress the activity of the ovaries and slow the growth of the endometrial tissue. Such drugs include but are not limited to oral contraceptives, progestins, danazol and GnRH (gonadotropin-releasing hormone) agonists.
Administration of the prenyl-protein transferase inhibitor may also be combined with surgical treatment of endometriosis (such as surgical removal of misplaced endometrial tissue) where appropriate.
The instant compounds may also be useful as inhibitors of corneal inflammation. These compounds may improve the treatment of corneal opacity which results from cauterization-induced corneal inflammation. The instant compounds may also be useful in reducing corneal edema and neovascularization. (K. Sonoda et al., Invest. Ophthalmol. Vis. Sci., 1998, vol. 39, p 2245-2251).
The compounds of this invention may be administered to mammals, preferably humans, either alone or, preferably, in combination with pharmaceutically acceptable carriers, excipients or diluents, in a pharmaceutical composition, according to standard pharmaceutical practice. The compounds can be administered orally or parenterally, including the intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration.
Additionally, the compounds of the instant invention may be administered to a mammal in need thereof using a gel extrusion mechanism (GEM) device, such as that described in U.S. Ser. No. 60/144,643, filed on Jul. 20, 1999, which is hereby incorporated by reference.
As used herein, the term xe2x80x9ccompositionxe2x80x9d is intended to encompass a product comprising the specified ingredients in the specific amounts, as well as any product which results, directly or indirectly, from combination of the specific ingredients in the specified amounts.
The pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may be for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, microcrystalline cellulose, sodium crosscarmellose, corn starch, or alginic acid; binding agents, for example starch, gelatin, polyvinyl-pyrrolidone or acacia, and lubricating agents, for example, magnesium stearate, stearic acid or talc. The tablets may be uncoated or they may be coated by known techniques to mask the unpleasant taste of the drug or delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a water soluble taste masking material such as hydroxypropyl-methylcellulose or hydroxypropylcellulose, or a time delay material such as ethyl cellulose, cellulose acetate buryrate may be employed.
Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water soluble carrier such as polyethyleneglycol or an oil medium, for example peanut oil, liquid paraffin, or olive oil.
Aqueous suspensions contain the active material in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethyl-cellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents may be a naturally-occurring phosphatide, for example lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethylene-oxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose, saccharin or aspartame.
Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and flavoring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as butylated hydroxyanisol or alpha-tocopherol.
Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, may also be present. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.
The pharmaceutical compositions of the invention may also be in the form of an oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or arachis oil, or a mineral oil, for example liquid paraffin or mixtures of these. Suitable emulsifying agents may be naturally-occurring phosphatides, for example soy bean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavoring agents, preservatives and antioxidants.
Syrups and elixirs may be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative, flavoring and coloring agents and antioxidant.
The pharmaceutical compositions may be in the form of a sterile injectable aqueous solutions. Among the acceptable vehicles and solvents that may be employed are water, Ringer""s solution and isotonic sodium chloride solution.
The sterile injectable preparation may also be a sterile injectable oil-in-water microemulsion where the active ingredient is dissolved in the oily phase. For example, the active ingredient may be first dissolved in a mixture of soybean oil and lecithin. The oil solution then introduced into a water and glycerol mixture and processed to form a microemulation.
The injectable solutions or microemulsions may be introduced into a patient""s blood-stream by local bolus injection. Alternatively, it may be advantageous to administer the solution or microemulsion in such a way as to maintain a constant circulating concentration of the instant compound. In order to maintain such a constant concentration, a continuous intravenous delivery device may be utilized. An example of such a device is the Deltec CADD-PLUS(trademark) model 5400 intravenous pump.
The pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension for intramuscular and subcutaneous administration. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example as a solution in 1,3-butane diol. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.
Compounds of Formula A may also be administered in the form of a suppositories for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol.
For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compound of Formula A are employed. (For purposes of this application, topical application shall include mouth washes and gargles.) The compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles and delivery devices, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in the art. To be administered in the form of a transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen. Compounds of the present invention may also be delivered as a suppository employing bases such as cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol.
When a compound according to this invention is administered into a human subject, the daily dosage will normally be determined by the prescribing physician with the dosage generally varying according to the age, weight, sex and response of the individual patient, as well as the severity of the patient""s symptoms.
In one exemplary application, a suitable amount of compound is administered to a mammal undergoing treatment for cancer. Administration occurs in an amount between about 0.1 mg/kg of body weight to about 60 mg/kg of body weight per day, preferably of between 0.5 mg/kg of body weight to about 40 mg/kg of body weight per day.
The compounds of the instant invention may also be co-administered with other well known therapeutic agents that are selected for their particular usefulness against the condition that is being treated. For example, the compounds of the instant invention may also be co-administered with other well known cancer therapeutic agents that are selected for their particular usefulness against the condition that is being treated. Included in such combinations of therapeutic agents are combinations of the instant prenyl-protein transferase inhibitors and an antineoplastic agent. It is also understood that such a combination of antineoplastic agent and inhibitor of prenyl-protein transferase may be used in conjunction with other methods of treating cancer and/or tumors, including radiation therapy and surgery. It is further understood that any of the therapeutic agents described herein may also be used in combination with a compound of the instant invention and an antineoplastic agent.
Examples of an antineoplastic agent include, in general, microtubule-stabilizing agents such as paclitaxel (also known as Taxol(copyright)), docetaxel (also known as Taxotere(copyright)), epothilone A, epothilone B, desoxyepothilone A, desoxyepothilone B or their derivatives); microtubule-disruptor agents; alkylating agents, for example, nitrogen mustards, ethyleneimine compounds, alkyl sulfonates and other compounds with an alkylating action such as nitrosoureas, cisplatin, and dacarbazine; anti-metabolites, for example, folic acid, purine or pyrimidine antagonists; epidophyllotoxin; an antineoplastic enzyme; a topoisomerase inhibitor; procarbazine; mitoxantrone; platinum coordination complexes; biological response modifiers and growth inhibitors; mitotic inhibitors, for example, vinca alkaloids and derivatives of podophyllotoxin; cytotoxic antibiotics; hormonal/anti-hormonal therapeutic agents, haematopoietic growth factors and antibodies (such as trastuzumab, also known as Herceptin(trademark)).
Example classes of antineoplastic agents include, for example, the anthracycline family of drugs, the vinca drugs, the mitomycins, the bleomycins, the cytotoxic nucleosides, the taxanes, the epothilones, discodermolide, the pteridine family of drugs, diynenes and the podophyllotoxins. Particularly useful members of those classes include, for example, doxorubicin, carminomycin, daunorubicin, aminopterin, methotrexate, methopterin, dichloro-methotrexate, mitomycin C, porfiromycin, 5-fluorouracil, 6-mercaptopurine, gemcitabine, cytosine arabinoside, podophyllotoxin or podo-phyllotoxin derivatives such as etoposide, etoposide phosphate or teniposide, melphalan, vinblastine, vincristine, leurosidine, vindesine, leurosine, paclitaxel and the like. Other useful antineoplastic agents include estramustine, cisplatin, carboplatin, cyclophosphamide, bleomycin, tamoxifen, ifosamide, melphalan, hexamethyl melamine, thiotepa, cytarabin, idatrexate, trimetrexate, dacarbazine, L-asparaginase, dactinomycin, mechlorethamine (nitrogen mustard), streptozocin, cyclophosphamide, carmustine (BCNU), lomustine (CCNU), procarbazine, mitomycin, cytarabine, etoposide, methotrexate, bleomycin, chlorambucil, camptothecin, CPT-11, topotecan, ara-C, bicalutamide, flutamide, leuprolide, pyridobenzoindole derivatives, interferons and interleukins. Particular examples of antineoplastic, or chemotherapeutic, agents are described, for example, by D. J. Stewart in xe2x80x9cNausea and Vomiting: Recent Research and Clinical Advancesxe2x80x9d, Eds. J. Kucharczyk, et al., CRC Press Inc., Boca Raton, Fla., USA (1991), pages 177-203, especially page 188. See also, R. J. Gralla, et al., Cancer Treatment Reports, 68(1), 163-172 (1984).
The preferred class of antineoplastic agents is the taxanes and the preferred antineoplastic agent is paclitaxel.
The compounds of the instant invention may also be co-administered with antisense oligonucleotides which are specifically hybridizable with RNA or DNA deriving from human ras gene. Such antisense oligonucleotides are described in U.S. Pat. No. 5,576,208 and PCT Publ. No. WO 99/22772. The instant compounds are particularly useful when co-administered with the antisense oligonucleotide comprising the amino acid sequence of SEQ.ID.NO: 2 of U.S. Pat. No. 5,576,208.
Certain compounds of the instant invention may exhibit very low plasma concentrations and significant inter-individual variation in the plasma levels of the compound. It is believed that very low plasma concentrations and high intersubject variability achieved following administration of certain prenyl-protein transferase inhibitors to mammals may be due to extensive metabolism by cytochrome P450 enzymes prior to entry of drug into the systemic circulation. Prenyl-protein transferase inhibitors may be metabolized by cytochrome P450 enzyme systems, such as CYP3A4, CYP2D6, CYP2C9, CYP2C19 or other cytochrome P450 isoform. If a compound of the instant invention demonstrates an affinity for one or more of the cytochrome P450 enzyme systems, another compound with a higher affinity for the P450 enzyme(s) involved in metabolism should be administered concomitantly. Examples of compounds that have a comparatively very high affinity for CYP3A4, CYP2D6, CYP2C9, CYP2C19 or other P450 isoform include, but are not limited to, piperonyl butoxide, troleandomycin, erythromycin, proadifen, isoniazid, allylisopropylacetamide, ethinylestradiol, chloramphenicol, 2-ethynylnaphthalene and the like. Such a high affinity compound, when employed in combination with a compound of formula A, may reduce the inter-individual variation and increase the plasma concentration of a compound of formula A to a level having substantial therapeutic activity by inhibiting the metabolism of the compound of formula A. Additionally, inhibiting the metabolism of a compound of the instant invention prolongs the pharmacokinetic half-life, and thus the pharmacodynamic effect, of the compound.
A compound of the present invention may be employed in conjunction with antiemetic agents to treat nausea or emesis, including acute, delayed, late-phase, and anticipatory emesis, which may result from the use of a compound of the present invention, alone or with radiation therapy. For the prevention or treatment of emesis a compound of the present invention may be used in conjunction with other anti-emetic agents, especially neurokinin-1 receptor antagonists, 5HT3 receptor antagonists, such as ondansetron, granisetron, tropisetron, and zatisetron, GABAB receptor agonists, such as baclofen, or a corticosteroid such as Decadron (dexamethasone), Kenalog, Aristocort, Nasalide, Preferid, Benecorten or others such as disclosed in U.S. Pat. Nos. 2,789,118, 2,990,401, 3,048,581, 3,126,375, 3,929,768, 3,996,359, 3,928,326 and 3,749,712. For the treatment or prevention of emesis, conjunctive therapy with a neurokinin-1 receptor antagonist, a 5HT3 receptor antagonist and a corticosteroid is preferred.
Neurokinin-1 receptor antagonists of use in conjunction with the compounds of the present invention are fully described, for example, in U.S. Pat. Nos. 5,162,339, 5,232,929, 5,242,930, 5,373,003, 5,387,595, 5,459,270, 5,494,926, 5,496,833, 5,637,699, 5,719,147; European Patent Publication Nos. EP 0 360 390, 0 394 989, 0 428 434, 0 429 366, 0 430 771, 0 436 334, 0 443 132, 0 482 539, 0 498 069, 0 499 313, 0 512 901, 0 512 902, 0 514 273, 0 514 274, 0 514 275, 0 514 276, 0 515 681, 0 517 589, 0 520 555, 0 522 808, 0 528 495, 0 532 456, 0 533 280, 0 536 817, 0 545 478, 0 558 156, 0 577 394, 0 585 913,0 590 152, 0 599 538, 0 610 793, 0 634 402, 0 686 629, 0 693 489, 0 694 535, 0 699 655, 0 699 674, 0 707 006, 0 708 101, 0 709 375, 0 709 376, 0 714 891, 0 723 959, 0 733 632 and 0 776 893; PCT International Patent Publication Nos. WO 90/05525, 90/05729, 91/09844, 91/18899, 92/01688, 92/06079, 92/12151, 92/15585, 92/17449, 92/20661, 92/20676, 92/21677, 92/22569, 93/00330, 93/00331, 93/01159, 93/01165, 93/01169, 93/01170, 93/06099, 93/09116, 93/10073, 93/14084, 93/14113, 93/18023, 93/19064, 93/21155, 93/21181, 93/23380, 93/24465, 94/00440, 94/01402, 94/02461, 94/02595, 94/03429, 94/03445, 94/04494, 94/04496, 94/05625, 94/07843, 94/08997, 94/10165, 94/10167, 94/10168, 94/10170, 94/11368, 94/13639, 94/13663, 94/14767, 94/15903, 94/19320, 94/19323, 94/20500, 94/26735, 94/26740, 94/29309, 95/02595, 95/04040, 95/04042, 95/06645, 95/07886, 95/07908, 95/08549, 95/11880, 95/14017, 95/15311, 95/16679, 95/17382, 95/18124, 95/18129, 95/19344, 95/20575, 95/21819, 95/22525, 95/23798, 95/26338, 95/28418, 95/30674, 95/30687, 95/33744, 96/05181, 96/05193, 96/05203, 96/06094, 96/07649, 96/10562, 96/16939, 96/18643, 96/20197, 96/21661, 96/29304, 96/29317, 96/29326, 96/29328, 96/31214, 96/32385, 96/37489, 97/01553, 97/01554, 97/03066, 97/08144, 97/14671, 97/17362, 97/18206, 97/19084, 97/19942 and 97/21702; and in British Patent Publication Nos. 2 266 529, 2 268 931, 2 269 170, 2 269 590, 2 271 774, 2 292 144, 2 293 168, 2 293 169, and 2 302 689. The preparation of such compounds is fully described in the aforementioned patents and publications.
A particularly preferred neurokinin-1 receptor antagonist for use in conjunction with the compounds of the present invention is 2-(R)-(1-(R)-(3,5-bis(trifluoromethyl)phenyl)ethoxy)-3-(S)-(4-fluorophenyl)-4-(3-(5-oxo-1H,4H-1,2,4-triazolo)methyl)morpholine, or a pharmaceutically acceptable salt thereof, which is described in U.S. Pat. No. 5,719,147.
For the treatment of cancer, it may be desirable to employ a compound of the present invention in conjunction with another pharmacologically active agent(s). A compound of the present invention and the other pharmacologically active agent(s) may be administered to a patient simultaneously, sequentially or in combination. For example, the present compound may employed directly in combination with the other active agent(s), or it may be administered prior, concurrent or subsequent to the administration of the other active agent(s). In general, the currently available dosage forms of the known therapeutic agents for use in such combinations will be suitable.
For example, a compound of the present invention may be presented together with another therapeutic agent in a combined preparation, such as with an antiemetic agent for simultaneous, separate, or sequential use in the relief of emesis associated with employing a compound of the present invention and radiation therapy. Such combined preparations may be, for example, in the form of a twin pack. A preferred combination comprises a compound of the present invention with antiemetic agents, as described above.
Radiation therapy, including x-rays or gamma rays which are delivered from either an externally applied beam or by implantation of tiny radioactive sources, may also be used in combination with the instant inhibitor of prenyl-protein transferase alone to treat cancer.
Additionally, compounds of the instant invention may also be useful as radiation sensitizers, as described in WO 97/38697, published on Oct. 23, 1997, and herein incorporated by reference.
The instant compounds may also be useful in combination with other inhibitors of parts of the signaling pathway that links cell surface growth factor receptors to nuclear signals initiating cellular proliferation. Thus, the instant compounds may be utilized in combination with farnesyl pyrophosphate competitive inhibitors of the activity of farnesyl-protein transferase or in combination with a compound which has Raf antagonist activity. The instant compounds may also be co-administered with compounds that are selective inhibitors of geranylgeranyl protein transferase.
In particular, if the compound of the instant invention is a selective inhibitor of farnesyl-protein transferase, co-administration with a compound(s) that is a selective inhibitor of geranylgeranyl protein transferase may provide an improved therapeutic effect.
In particular, the compounds disclosed in the following patents and publications may be useful as farnesyl pyrophosphate-competitive inhibitor component of the instant composition: U.S. Ser. Nos. 08/254,228 and 08/435,047. Those patents and publications are incorporated herein by reference.
In practicing methods of this invention, which comprise administering, simultaneously or sequentially or in any order, two or more of a protein substrate-competitive inhibitor and a farnesyl pyrophosphate-competitive inhibitor, such administration can be orally or parenterally, including intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and topical routes of administration. It is preferred that such administration be orally. It is more preferred that such administration be orally and simultaneously. When the protein substrate-competitive inhibitor and farnesyl pyrophosphate-competitive inhibitor are administered sequentially, the administration of each can be by the same method or by different methods.
The instant compounds may also be useful in combination with an integrin antagonist for the treatment of cancer, as described in U.S. Ser. No. 09/055,487, filed Apr. 6, 1998, and WO 98/44797, published on Oct. 15, 1998, which are incorporated herein by reference.
As used herein the term an integrin antagonist refers to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to an integrin(s) that is involved in the regulation of angiogenisis, or in the growth and invasiveness of tumor cells. In particular, the term refers to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to the xcex1vxcex23 integrin, which selectively antagonize, inhibit or counteract binding of a physiological ligand to the xcex1vxcex25 integrin, which antagonize, inhibit or counteract binding of a physiological ligand to both the xcex1vxcex23 integrin and the xcex1vxcex25 integrin, or which antagonize, inhibit or counteract the activity of the particular integrin(s) expressed on capillary endothelial cells. The term also refers to antagonists of the xcex11xcex21, xcex12xcex21, xcex15xcex21, xcex16xcex21 and xcex16xcex24 integrins. The term also refers to antagonists of any combination of xcex1vxcex23 integrin, xcex1vxcex25 integrin, xcex11xcex21, xcex11xcex21, xcex12xcex21, xcex15xcex21, xcex16xcex21 and xcex16xcex24 integrins. The instant compounds may also be useful with other agents that inhibit angiogenisis and thereby inhibit the growth and invasiveness of tumor cells, including, but not limited to angiostatin and endostatin.
The instant compounds may also be useful in combination with an inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase) for the treatment of cancer. Compounds which have inhibitory activity for HMG-CoA reductase can be readily identified by using assays well-known in the art. For example, see the assays described or cited in U.S. Pat. No. 4,231,938 at col. 6, and WO 84/02131 at pp. 30-33. The terms xe2x80x9cHMG-CoA reductase inhibitorxe2x80x9d and xe2x80x9cinhibitor of HMG-CoA reductasexe2x80x9d have the same meaning when used herein.
Examples of HMG-CoA reductase inhibitors that may be used include but are not limited to lovastatin (MEVACOR(copyright); see U.S. Pat. Nos. 4,231,938; 4,294,926; 4,319,039), simvastatin (ZOCOR(copyright); see U.S. Pat. Nos. 4,444,784; 4,820,850; 4,916,239), pravastatin (PRAVACHOL(copyright); see U.S. Pat. Nos. 4,346,227; 4,537,859; 4,410,629; 5,030,447 and 5,180,589), fluvastatin (LESCOL(copyright); see U.S. Pat. Nos. 5,354,772; 4,911,165; 4,929,437; 5,189,164; 5,118,853; 5,290,946; 5,356,896), atorvastatin (LIPITOR(copyright); see U.S. Pat. Nos. 5,273,995; 4,681,893; 5,489,691; 5,342,952) and cerivastatin (also known as rivastatin and BAYCHOL(copyright); see U.S. Pat. No. 5,177,080). The structural formulas of these and additional HMG-CoA reductase inhibitors that may be used in the instant methods are described at page 87 of M. Yalpani, xe2x80x9cCholesterol Lowering Drugsxe2x80x9d, Chemistry and Industry, pp. 85-89 (Feb. 5, 1996) and U.S. Pat. Nos. 4,782,084 and 4,885,314. The term HMG-CoA reductase inhibitor as used herein includes all pharmaceutically acceptable lactone and open-acid forms (i.e., where the lactone ring is opened to form the free acid) as well as salt and ester forms of compounds which have HMG-CoA reductase inhibitory activity, and therefor the use of such salts, esters, open-acid and lactone forms is included within the scope of this invention. An illustration of the lactone portion and its corresponding open-acid form is shown below as structures I and II. 
In HMG-CoA reductase inhibitors where an open-acid form can exist, salt and ester forms may preferably be formed from the open-acid, and all such forms are included within the meaning of the term xe2x80x9cHMG-CoA reductase inhibitorxe2x80x9d as used herein. Preferably, the HMG-CoA reductase inhibitor is selected from lovastatin and simvastatin, and most preferably simvastatin. Herein, the term xe2x80x9cpharmaceutically acceptable saltsxe2x80x9d with respect to the HMG-CoA reductase inhibitor shall mean non-toxic salts of the compounds employed in this invention which are generally prepared by reacting the free acid with a suitable organic or inorganic base, particularly those formed from cations such as sodium, potassium, aluminum, calcium, lithium, magnesium, zinc and tetramethylammonium, as well as those salts formed from amines such as ammonia, ethylenediamine, N-methylglucamine, lysine, arginine, ornithine, choline, N,Nxe2x80x2-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, 1-p-chlorobenzyl-2-pyrrolidine-1xe2x80x2-yl-methylbenzimidazole, diethylamine, piperazine, and tris(hydroxymethyl) aminomethane. Further examples of salt forms of HMG-CoA reductase inhibitors may include, but are not limited to, acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynapthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylsulfate, mucate, napsylate, nitrate, oleate, oxalate, pamaote, palmitate, panthothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate.
Ester derivatives of the described HMG-CoA reductase inhibitor compounds may act as prodrugs which, when absorbed into the bloodstream of a warm-blooded animal, may cleave in such a manner as to release the drug form and permit the drug to afford improved therapeutic efficacy.
Similarly, the instant compounds may be useful in combination with agents that are effective in the treatment and prevention of NF-1, restenosis, polycystic kidney disease, infections of hepatitis delta and related viruses and fungal infections.
If formulated as a fixed dose, such combination products employ the combinations of this invention within the dosage range described above and the other pharmaceutically active agent(s) within its approved dosage range. Combinations of the instant invention may alternatively be used sequentially with known pharmaceutically acceptable agent(s) when a multiple combination formulation is inappropriate.
The instant compounds may also be useful in combination with prodrugs of antineoplastic agents. In particular, the instant compounds may be co-administered either concurrently or sequentially with a conjugate (termed a xe2x80x9cPSA conjugatexe2x80x9d) which comprises an oligopeptide, that is selectively cleaved by enzymatically active prostate specific antigen (PSA), and an antineoplastic agent. Such co-administration will be particularly useful in the treatment of prostate cancer or other cancers which are characterized by the presence of enzymatically active PSA in the immediate surrounding cancer cells, which is secreted by the cancer cells.
Compounds which are PSA conjugates and are therefore useful in such a co-administration, and methods of synthesis thereof, can be found in the following patents, pending patent applications and publications which are herein incorporated by reference:
U.S. Pat. No. 5,599,686, granted on Feb. 4, 1997;
WO 96/00503 (Jan. 11, 1996); U.S. Ser. No. 08/404,833, filed on Mar. 15, 1995;
U.S. Ser. No. 08/468,161, filed on Jun. 6, 1995;
U.S. Pat. No. 5,866,679, granted on Feb. 2, 1999;
WO 98/10651 (Mar. 19, 1998); U.S. Ser. No. 08/926,412, filed on Sep. 9, 1997;
WO 98/18493 (May 7, 1998); U.S. Ser. No. 08/950,805, filed on Oct. 14, 1997;
WO 99/02175 (Jan. 21, 1999); U.S. Ser. No. 09/112,656, filed on Jul. 9, 1998; and
WO 99/28345 (Jun. 10, 1999); U.S. Ser. No. 09/193,365, filed on Nov. 17, 1998.
Compounds which are described as prodrugs wherein the active therapeutic agent is released by the action of enzymatically active PSA and therefore may be useful in such a co-administration, and methods of synthesis thereof, can be found in the following patents, pending patent applications and publications, which are herein incorporated by reference: WO 98/52966 (Nov. 26, 1998).
All patents, publications and pending patent applications identified are herein incorporated by reference.
The compounds of the instant invention are also useful as a component in an assay to rapidly determine the presence and quantity of farnesyl-protein transferase (FPTase) in a composition. Thus the composition to be tested may be divided and the two portions contacted with mixtures which comprise a known substrate of FPTase (for example a tetrapeptide having a cysteine at the amine terminus) and farnesyl pyrophosphate and, in one of the mixtures, a compound of the instant invention. After the assay mixtures are incubated for an sufficient period of time, well known in the art, to allow the FPTase to farnesylate the substrate, the chemical content of the assay mixtures may be determined by well known immunological, radiochemical or chromatographic techniques. Because the compounds of the instant invention are selective inhibitors of FPTase, absence or quantitative reduction of the amount of substrate in the assay mixture without the compound of the instant invention relative to the presence of the unchanged substrate in the assay containing the instant compound is indicative of the presence of FPTase in the composition to be tested.